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. 2024 Sep 16;15:1458967. doi: 10.3389/fimmu.2024.1458967

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Copyright © 2024 Govaerts, Van Breedam, De Beuckeleer, Goethals, D’Incal, Di Stefano, Van Calster, Buyle-Huybrecht, Boeren, De Reu, Paludan, Thiry, Lebrun, Sadzot-Delvaux, Motaln, Rogelj, Van Weyenbergh, De Vos, Vanden Berghe, Ogunjimi, Delputte and Ponsaerts

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Immune responsiveness of VZV^eGFP-ORF23- and SeV^eGFP-infected NSPHs. (A) Representative live cell fluorescence image of 5-month-old control NSPHs (CTRL), NSPHs inoculated with eGFP+ control ARPE19 cells (ARPE-CTRL, green), NSPHs inoculated with VZV^eGFP-ORF23-infected ARPE19 cells (VZV^eGFP-ORF23, green), and NSPHs infected with SeV^eGFP (SeV^eGFP, green) at day 7. Scale bars of 50 and 200 µm are indicated on the images. (B) Representative images of 5-month-old control NSPHs (CTRL), NSPHs inoculated with eGFP+ control ARPE19 cells (ARPE-CTRL, green), NSPHs inoculated with VZV^eGFP-ORF23-infected ARPE19 cells (VZV^eGFP-ORF23, green), and NSPHs infected with SeV^eGFP (SeV^eGFP, green) at day 7. Nuclei are labelled with DAPI (blue). Scale bars of 50 and 200 µm are indicated on the images. (C) Boxplots showing IL-6 and CXCL10 cytokine secretion (in pg/mL) by 5-month-old control NSPHs (CTRL, n=4) and NSPHs stimulated with IL-1b, TNF-α, IFN-γ, LPS and ATP (n=4) at day 3 post-stimulation (dps). **** p<0,0001. Error bars indicate standard deviation (SD). (D) Combined box/dotplots showing IL-6 and CXCL10 cytokine secretion (in pg/mL) derived from 2 independent experiments for5-month-old control NSPHs (CTRL, n=8 for Exp1, n=4 for Exp 2), NSPHs inoculated with eGFP+ control ARPE19 cells (ARPE-CTRL, n=8 for Exp1, n=4 for Exp 2), NSPHs inoculated with VZV^eGFP-ORF23-infected ARPE19 cells (VZV, n=8 for Exp1, n=4 for Exp 2), and NSPHs infected with SeV^eGFP (SeV, n=8 for Exp1, n=4 for Exp 2) at 3-, 5- and 7-days post-infection (dpi). The mean of each individual experiment is given as a dot within the boxplot. ** p<0,01. **** p<0,0001.